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Attain the optimal resolution of mono- and disaccharides using the Thermo Scientific™ Dionex™ CarboPac™ SA10-4µm column. Compared to the previous Dionex CarboPac SA10 column, the new column uses smaller resin particles for more efficient separations resulting in more accurate peak integration and more reliable results. The Dionex CarboPac SA10-Fast-4µm columns are shorter anion-exchange columns for the fast separation and accurate quantification of mono- and disaccharides in biofuels, foods and beverages. The column provides:

• Fast analysis and short separation times

• High resolution

• High capacity

• High reproducibility

• Superior peak efficiency

Thermo Scientific Dionex CarboPac SA10-4µm Column for Fast, High-Resolution Mono- and Disaccharide Analysis

Fast, High-Resolution Determination of Mono- and Disaccharides The Dionex CarboPac SA10-Fast-4µm anion-exchange column is optimized for the fast chromatographic analysis of biofuels and foods for simple sugars. The fast, efficient separations of mono- and disaccharides are achieved without compromising resolution. For the eight most common biofuel mono- and disaccharides, the Dionex CarboPac SA10-Fast-4µm column provides the unsurpassed fast separation time of less than 5 min. The seven most common food sugars can be separated in the exceptionally fast time of less than 6 min. Due to the high capacity of the Dionex CarboPac SA10-4µm column, minimum sample dilution is required.

The Dionex CarboPac SA10-4µm columns are composed of a unique 4 µm supermacroporous (SMP) substrate coated with a strong anion-exchange layer of latex nano beads, which provides high capacity and rapid, high-resolution separations. Excellent resolution and calibration linearity results in accurate quantitative carbohydrate analysis. The Dionex CarboPac SA10-4µm anion-exchange column is used with pulsed amperometric detection (PAD) systems and permits the direct quantification of nonderivatized carbohydrates with minimal sample preparation. The column provides high reproducibility, and 1000 injection cycles under high pH conditions are possible. The Dionex CarboPac SA10-4µm column is available in microbore (2 mm) and standard bore (4 mm) formats.

Separation of Biofuel Sugars Using the Dionex CarboPac SA10-Fast-4µm Column

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Biofuel and Food ApplicationsThe biofuel, food, and beverage industries process very high volumes of carbohydrate samples, and require high throughput mono- and di-saccharide analysis capabilities. The Dionex CarboPac SA10-4µm column is specifically designed for the exceptionally fast, high-resolution analysis of carbohydrate samples for biofuel and food applications. Rapid separation of the eight most common biofuel mono- and disaccharides can be achieved on the (a) CarboPac SA10-Fast-4μm (2 × 150 mm) column in less than 5 minutes and (b) the Dionex CarboPac SA10-4μm (2 × 250 mm) column in less than 6 minutes (Figure 1). The capability to analyze the eight most common food sugars with a short separation time is important for many applications. The Dionex CarboPac SA10-Fast-4µm column can separate these seven common food sugars in less than 8 min, as shown in Figure 2(a).

High Concentration Samples The Dionex CarboPac SA10-4µm column is capable of analyzing a broad range of sample concentrations, including high-concentration samples. Packed with highly porous resin beads, the Dionex CarboPac SA10-4µm column has significantly higher capacity than other Dionex CarboPac columns and is the column of choice for high-concentration mono- and disaccharide sample analysis. When high-concentration simple sugar samples are separated on a Dionex CarboPac SA10-4µm column, linearity is maintained and accurate quantitative analysis is achieved. Figure 3 demonstrates the separation of a high-concentration corn stover hydrolysate sample on the Dionex CarboPac SA10-4µm 2 mm column. Note excellent separation of biofuel sugars when 43 g/L corn stover hydrolysate sample was injected using 1:200 dilution of the sample and optimum gasket thickness (15 mil for the 2 mm column or 62 mil for the 4 mm column).

Figure 1. Fast separation of biofuel sugars on the (a) Dionex CarboPac SA10-Fast-4µm (2 × 150 mm) and (b) Dionex CarboPac SA10-4µm (2 × 250 mm)

Figure 2. Fast separation of food sugars on the (a) Dionex CarboPac SA10-Fast-4µm (2 × 150 mm) and (b) Dionex CarboPac SA10-4µm (2 × 250 mm)

Column: See chromatogram Eluent: 1 mM KOHEluent Source: Dionex EGC 500 KOH cartridgeFlow Rate: 0.38 mL/minInjection Volume: 2.5 µLGasket: 2 milTemperature: 45 °CDetection: Integrated Amperometry, quadruple pulse waveformWorking Electrode: PTFE Gold, disposable electrodeReference Electrode: Silver/Silver ChlorideStandard: 10 mg/L

Peaks: 1. Fucose 2. Sucrose 3. Arabinose 4. Galactose 5. Glucose 6. Xylose 7. Mannose 8. Fructose

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Column: See chromatogram Eluent: 12 mM KOHEluent Source: Dionex EGC 500 KOH cartridgeFlow Rate: 0.38 mL/minInj. Volume: 2.5 µLTemperature: 40 °CDetection: Integrated Amperometry, quadruple pulse waveformWorking Electrode: PTFE Gold, disposable electrodeReference Electrode: Silver/Silver ChlorideSample: 5 mg/L each

Peaks: 1. Sucrose 2. Glucose 3. Fructose 4. Lactose 5. Cellibiose 6. Maltose

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Figure 3. High-resolution separation of a high concentration corn stover hydrolysate sample using a Dionex CarboPac SA10-4µm 2 mm column and 15 mil gasket thickness.

Column: Dionex CarboPac SA10-4µm Guard and Analytical columns, 2 mmEluent: 1 mM KOHEluent Source: Dionex EGC 500 KOH cartridgeFlow Rate: 0.38 mL/minInj. Volume: 2.5 µLGasket: 15 milTemperature: 45 °CDetection: Integrated Amperometry, quadruple pulse waveformWorking Electrode: PTFE Gold, disposable electrodeReference Electrode: Silver/Silver ChlorideSample: Corn Stover 43 g/L, 1:200 Dilution

Peaks: 1. Arabinose 2. Galactose 3. Glucose 4. Xylose 5. Fructose

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System CompatibilityThe Dionex CarboPac SA10-4µm column is compatible with KOH eluents prepared by Eluent Generation only. It is not compatible with manually prepared eluents. The smaller particles used in the Dionex CarboPac SA10-4μm column generate higher backpressure under standard operating conditions and therefore cannot can be used with Thermo Scientific Dionex IC systems that are not high-pressure compatible. A Thermo Scientific™ Dionex™ Reagent-Free™ HPIC System equipped with an Eluent Generator and Electrochemical Detector, such as the Thermo Scientific™ Dionex™ ICS-5000+ HPIC™ System, is required.

Innovative Resin TechnologyThe Dionex CarboPac SA10-4µm column resin consists of 4 μm diameter super macroporous resin beads covered with the quaternary ammonium ion functionalized nano beads. This wide-pore resin structure permits fast mass transfer, resulting in high-resolution chromatography. The highly porous beads have large surface areas, providing high capacity. Together, the morphology of the porous substrate and agglomerated functionalized nano beads results in fast, high-resolution separations of common sugars with exceptionally short analysis times. The smaller particles used in the Dionex CarboPac SA10-4µm column result in excellent peak efficiencies and higher resolution separations compared to previous generation columns, see Figure 4. Improved resolution makes peak integration easier and more reliable leading to more accurate results.

Figure 4. Comparison of Dionex (a) CarboPac SA10-Fast-4 μm (2 mm), (b) CarboPac SA10-4μm (2 mm), and (c) CarboPac SA10 (2 mm) for the

analysis of 8 Biofuel sugars.

Column: See chromatogramEluent: 1 mM KOHEluent Source: Dionex EGC 500 KOH cartridgeFlow Rate: 0.38 mL/minInj. Volume: 2.5 µLTemperature: 45 °CDetection: Integrated Amperometry, quadruple pulse waveformWorking Electrode: PTFE Gold, disposable electrodeReference Electrode: Silver/Silver ChlorideStandard: 10 mg/L each

Peaks: 1. Fucose 2. Sucrose 3. Arabinose 4. Galactose 5. Glucose 6. Xylose 7. Mannose 8. Fructose

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(c) Dionex CarboPac SA10 ColumnColumn Pressure = 1240 psi

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(b) Dionex CarboPac SA10-4µm Column Column Pressure = 3415 psi

(a) Dionex CarboPac SA10-Fast-4µm Column Column Pressure = 2233 psi

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Dionex CarboPac SA10-4μm column, 2 mmDionex CarboPac SA10-Fast-4μm column, 2 mm Standard Dionex CarboPac SA10 column, 2 mm

No. Peak Name Ret.Time

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Resolution

1 Fucose 2.7 11602 6.13

2 Sucrose 3.5 9055 2.44

3 Arabinose 3.8 13729 2.05

4 Galactose 4.1 12105 2.84

5 Glucose 4.5 11377 3.15

6 Xylose 5.1 14128 1.73

7 Mannose 5.4 14337 1.95

8 Fructose 5.7 13416 n.a.

No. Peak Name Ret.Time

(min) Plates (EP)

Resolution

1 Fucose 1.8 7045 4.73

2 Sucrose 2.3 5602 1.95

3 Arabinose 2.5 8459 1.58

4 Galactose 2.7 7317 2.25

5 Glucose 3.0 7062 2.45

6 Xylose 3.4 8508 1.26

7 Mannose 3.6 8236 1.65

8 Fructose 3.8 7915 n.a.

No. Peak NameRet.Time

(min) Plates (EP)

Resolution

1 Fucose 2.7 7103 4.71

2 Sucrose 3.5 5391 1.84

3 Arabinose 3.8 7706 1.53

4 Galactose 4.1 6862 2.07

5 Glucose 4.5 6720 2.37

6 Xylose 5.1 7803 1.21

7 Mannose 5.4 7901 1.63

8 Fructose 5.8 7211 n.a.

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Table 1. Peak area reproducibility: electrode-to-electrode for five disposable electrodes.

Electrode Average Fuc GalN GlcN Gal Glc Man

1 n = 12 3.09 7.05 5.58 4.92 5.82 4.22

2 n = 12 3.13 7.49 5.95 4.68 6.24 4.25

3 n = 12 3.35 7.65 6.04 5.32 6.23 4.52

4 n = 12 3.12 6.76 5.33 4.70 5.52 4.18

5 n = 12 3.12 6.93 5.46 4.83 5.54 4.20

Average 3.16 7.18 5.67 4.89 5.87 4.27

S.D 0.11 0.38 0.31 0.26 0.36 0.14

Elec-to-Elec RSD 3.41% 5.30% 5.44% 4.35% 6.05% 3.28%

Perm. Elec n = 12 2.91 5.79 4.36 4.84 4.51 3.64

Figure 5. Consistent separation results obtained after 1000 separations of a biofuel sugar sample using a Dionex CarboPac SA10-4µm column.

Column: Dionex CarboPac SA10-4µm Guard and Analytical columns, 2mmEluent: 1 mM KOHEluent Source: Dionex EGC 500 KOH cartridgeFlow Rate: 0.38 mL/minInj. Volume: 2.5 µLGasket: 2 milTemperature: 45 °CDetection: Integrated Amperometry, quadruple pulse waveformWorking Electrode: PTFE Gold, disposable electrodeReference Electrode: Silver/Silver ChlorideStandard: 10 mg/L

Peaks: 1. Fucose 2. Sucrose 3. Arabinose 4. Galactose

5. Glucose6. Xylose7. Mannose8. Fructose

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High ReproducibilityThe Dionex CarboPac SA10-4µm column provides consistent chromatographic performance and highly reproducible analysis. Figure 5 shows the high reproducibility of the column, with consistent separation results obtained after 1000 runs of a biofuel sugar sample. The Dionex CarboPac SA10-4µm column needs to be used with the on-line eluent generator, (EG), which produces carbonate-free hydroxide eluent to ensure reproducible separations.

Disposable Gold ElectrodesPulsed amperometric detection (PAD) permits detection of carbohydrates with excellent signal-to-noise ratios down to approximately 10 picomoles without requiring derivitization. Carbohydrates are detected by measuring the electrical current generated by their oxidation at the surface of a gold electrode. A gold electrode is commonly used for determination of monosaccharides and disaccharides.

Although carbohydrates can be oxidized at a gold working electrode, over time the products of oxidation reactions can foul the surface of the electrode, inhibiting analyte oxidation. Electrode fouling with oxidation byproducts or sample components will reduce response, requiring working electrode polishing to restore the surface. Thermo Scientific Dionex disposable gold electrodes eliminate the need for electrode reconditioning (i.e. polishing). Disposable electrodes are economical, and thus can be replaced frequently while delivering peak area reproducibility superior to conventional Au working electrodes. The peak area reproducibility for five disposable electrodes is shown in Table 1.

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Ordering InformationIn the U.S., call (800) 346-6390 or contact the Thermo Fisher Scientific Regional Office nearest you. Outside the U.S., order through your local Thermo Fisher Scientific office or distributor. Refer to the following part numbers.

Analytical and Guard Columns Part Number

Dionex CarboPac SA10-Fast-4µm Analytical Column (4 × 150 mm) 088652

Dionex CarboPac SA10-Fast-4µm Guard Column (4 × 30 mm) 088654

Dionex CarboPac SA10-Fast-4µm Analytical Column (2 × 150 mm) 088653

Dionex CarboPac SA10-Fast-4µm Guard Column (2 × 30 mm) 088655

Dionex CarboPac SA10-4µm Analytical Column (4 × 250 mm) 088233

Dionex CarboPac SA10-4µm Guard Column (4 × 50 mm) 088234

Dionex CarboPac SA10-4µm Analytical Column (2 × 250 mm) 088235

Dionex CarboPac SA10-4µm Guard Column (2 × 50 mm) 088236

Electrodes Part Number

Disposable Electrodes for Carbohydrates, Pack of six Au (Polyester), two 0.002" gaskets 060139

Disposable Au (PTFE) Electrodes for Carbohydrates, Pack of six Au (PTFE), four, 0.002" gaskets 066480

Polypropylene Support Block, ED cell for use with Disposable Electrodes 062158

Gaskets, (PTFE) for Disposable Electrode 0.001" Pack of two 072117

Gaskets, (PTFE) for Disposable Electrode 0.002" Pack of four 060141

Gaskets, (ULTEM) for Disposable Electrodes 0.001" Pack of four 069339

Gasket, (HDPE) for Disposable Electrodes 0.015" 057364

Gasket, (UHMW PE) for Disposable Electrodes, 0.062" 075499

Polypropylene Support Block, for use with 0.062" gasket, ED Cell, for Disposable Electrodes 075501

ED Kit with 62 mil gasket, Polypropylene Support Block, for use with Disposable Electrodes 075502

Polypropylene Support Block, ED50A/50/40 Cell for use with Disposable Electrodes 060297

PS70915-EN 0615M

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DIONEX CARBOPAC SA10 COLUMN SPECIFICATIONS

Resin Characteristics

Particle Size 4 μm supermacroporous

Pore Size 2000 Å

Substrate Ethylvinylbenzene crosslinked with divinylbenzene

Crosslinking 55%

Ion-Exchange Capacity 290 μeq per 4.0 × 250 mm column; 72.5 µeq per 2.0 × 250 mm column

Latex Nano Bead Characteristics

Functional Group Difunctional quaternary ammonium ion

Latex Diameter 55 nm

Latex Crosslinking 4.5%

Typical Operating Parameters

pH Range 0–14

Temperature Range 4–60 °C

Recommended Flow Rate 1.5 mL/min (4 × 250 mm column); 0.38 mL/min (2 × 250 mm column)

Flow Rate Range 1.0–2.0 mL/min (4 × 250 mm column); 0.25–0.50 mL/min (2 × 250 mm column)

Pressure Limit 5000 psi

Organic Solvent Limit 100% compatible

Typical Eluents Potassium hydroxide or sodium hydroxide from Eluent Generation